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In Vitro and In Vivo Primary Metabolic Characterization of F18, a Novel Histone Deacetylase-6 (HDAC6) Inhibitor, Using UHPLC–QqQ–MS/MS and Q-TOF–MS Methods

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Abstract

F18, N-hydroxy-4-(2-methoxy-5-(methyl (2-methylquinazolin-4-yl) amino) phenoxy) butanamide, is a novel selective HDAC6 inhibitor with good antitumor activity. In the early drug development, drug-metabolism studies are a crucial and indispensable part. In this study, we proposed to evaluate the in vitro primary metabolism of F18 in phase Ι in liver microsomes from human, rat, dog, monkey and mouse and investigate the metabolite profile both in vitro and in vivo using LC–MS/MS methods. F18 showed high metabolic stability in human, rat, dog, monkey and mouse liver microsomes over 120 min, with t 1/2 >8 h in human, rat, and dog, and t 1/2 <3.5 h in monkey, with almost no clearance in mouse. Human cytochrome P450 (P450) phenotyping showed that F18 was predominantly metabolized by CYP2C9, CYP2E1, CYP2D6 and CYP3A4. The investigation of the effect of F18 on CYP enzymes in HLM demonstrated that this compound did not significantly inhibit CYP 1A2 (IC50 >100 μM), was a moderate inhibitor of CYP3A4 (IC50 = 1.63 μM) and had negligible effects on CYP3A1/2 activity in rats. The results will be valuable in understanding drug–drug interactions (DDI) when F18 is co-administered with other drugs. The metabolites of F18 were investigated in rat plasma, urine, feces and different liver microsomes in NADPH samples, yielding at least 11 metabolites in these biological samples. The prominent metabolic pathways were de-methylation, de-amination, de-oxidation and O-glucuronidation. In summary, this work provides the first clues regarding F18 metabolism, providing important information for comprehensive understanding of F18 metabolites.

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Acknowledgments

The authors state no conflicts of interest. This work was supported by the National Key Programs of China during the 12th Five-Year Plan Period (2012ZX09103101-009) and Guangdong Innovative Research Team Program (no. 2011Y073).

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Authors and Affiliations

  1. State Key Laboratory of Biotherapy, Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Keyuan Road 4, Gaopeng Street, Chengdu, 610041, People’s Republic of China

    Xiaobin Li, Minghai Tang, Hairong Wang, Haoyu Ye, Chunyu Wang, Qiunan Yang & Lijuan Chen

  2. Key Laboratory of Standardization of Chinese Herbal Medicine, School of Pharmacy, Chengdu University of TCM, The Ministry of Education , Chengdu, 611137, People’s Republic of China

    Xiaobin Li, Hairong Wang, Qiunan Yang & Li Wan

  3. Division of Nephrology, Kidney Research Institute, West China Hospital, West China Medical School, Sichuan University, Chengdu, Sichuan, 610041, People’s Republic of China

    Liang Ma

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  1. Xiaobin Li
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  3. Hairong Wang
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Correspondence to Minghai Tang.

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Whole procedures involving animals had been approved by the Institute Guidelines on Animal Experimentation of Sichuan University in China.

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X. Li and M. Tang are joint first authors.

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Li, X., Tang, M., Wang, H. et al. In Vitro and In Vivo Primary Metabolic Characterization of F18, a Novel Histone Deacetylase-6 (HDAC6) Inhibitor, Using UHPLC–QqQ–MS/MS and Q-TOF–MS Methods. Chromatographia 79, 1479–1490 (2016). https://doi.org/10.1007/s10337-016-3163-4

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  • DOI: https://doi.org/10.1007/s10337-016-3163-4

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